COP9信号小体复合体基因缺失对新型隐球酵母菌生长与毒力效应的影响  

Influence of COP9 signalosome complex gene deletion on growth and virulence of Cryptococcus neoformans

作　　者：樊一斌[1] 潘卫利[1] 陶小华[1] 皇幼明[1] 徐珍[1] 

机构地区：[1]浙江省人民医院皮肤科,浙江杭州310014

出　　处：《中华医院感染学杂志》2016年第3期533-536,共4页Chinese Journal of Nosocomiology

基　　金：浙江省自然科学基金青年基金资助项目(Q14H190009)

摘　　要：目的研究COP9信号小体复合体基因缺失对新型隐球酵母菌生长与毒力效应的影响,为研究抗真菌药物作用靶点提供试验数据。方法 2014年4月以临床标准株H99为背景,分别敲除其中一个目前已知的COP9编码基因,构建COP9信号小体复合体基因缺失菌株,通过高温、氧化、NO、高盐、高渗、细胞膜、细胞壁体外应激应答反应;进一步检测生长速度,绘制生长曲线;并以巨噬细胞感染模型、蜡蛾感染模型检测新型隐球酵母菌活体内毒力变化。结果与标准株相比,CSN1201△缺失株对温度、氧化、NO、高盐、高渗、细胞壁及细胞膜应激反应的敏感性显著升高;CSN1201基因缺失对新型隐球酵母菌的生长速率有一定的影响,而且高温条件更加重了其影响;CSN1201△缺失株和CSN7△缺失株的胞内生存率显著下降(PCSN1201△=0.003,PCSN7△<0.001),而其他缺失株则与标准株类似;与标准株H99相比,CSN1201△缺失株和CSN7△缺失株的毒力显著下降,平均生存时间分别为(23.5±0.95)d和(21.2±1.28)d(P<0.05);而CSN4△则表现为毒力显著增强,平均生存时间为(6.4±0.36)d(P<0.05),其余缺失株的毒力与标准株比较,差异无统计学意义。结论 COP9信号小体复合体的基因,尤其是CSN1201对于新型隐球酵母菌的毒力有一定的影响,可能是调控多种应激条件下新型隐球酵母菌毒力的重要基因,有可能成为抗真菌药物靶点。OBJECTIVE To provide experimental data for research on the targets of antifungal drugs by studying the influence of COP9 signalosome complex gene deletion on growth and virulence of Cryptococcus neoformans.METHODS COP9 signalosome complex gene deletion strains were constructed by knocking out one of the known COP9 encoding gene of clinical standard strains H99 in April 2014.High temperature,oxidation,NO,high salt,high permeability,cell membrane and cell wall stress response in vitro were tested.And the growth speed was tested further,and the growth curve was drawn.Macrophage infection model and Galleria mellonella infection model were used to detect virulence changes of C.neoformans in vivo.RESULTS Compared with the standard strains,CSN1201△strain showed significantly increased sensitivity to temperature,oxidation,NO,high salt,high permeability,cell membrane and cell wall stress reaction.CSN1201 gene deletion had a certain influence on the growth rate of C.neoformans,and high temperature conditions could enhance the impact.The intracellular survival rate of CSN1201△strain and CSN7△strain decreased significantly（PCSN1201△=0.003,PCSN7△0.001）,while the other strains were similar to standard strains.Compared with the standard strain H99,the virulence of CSN1201 △strain and CSN7△strain showed a significant reduction,and the average survival time was（23.5±0.95）d and（21.2±1.28）d respectively（P〈0.05）.The virulence of CSN4 △strain was remarkably increased,and the average survival time was（6.4±0.36）d（P〈0.05）.The virulence of the rest strains had no significant difference compared with standard strains.CONCLUSIONCOP9 signalosome complex gene,especially CSN1201,has a certain influence on the virulence of C.neoformans,which may be the important gene to regulate C.neoformans under various stress conditions and could be the antifungal drug target.

关 键 词：新型隐球酵母菌 COP9信号小体 应激应答 胞内生存率 毒力 

分 类 号：R379[医药卫生—病原生物学]